骆驼刺根际细菌的植物促生能力

采用纯培养方法使用5种培养基分析沙漠干旱植物骆驼刺的根际可培养细菌群落,并用盆钵试验验证这些菌株的植物促生能力.共从骆驼刺根际土中分离纯化了120株细菌,根据16S rRNA基因序列划分成32个16S r RNA基因型,分布在Actinobacteria、Firmicutes、Proteobacteria和Bacteroidetes等4个门的17个属内.其中,Actinobacteria占全部分离菌株的77.3%,是骆驼刺根际的优势微生物;该门的Streptomyces和Arthrobacter两个属是分离菌株最多的属.从16个型(55株)的细菌中检测到了固氮酶nif H基因,占全部分离菌株的45.8%、全部型的50%.盆钵试验中,菌株Microbacterium sp.WLJ053、Streptomyces sp.WLJ079、Paenibacillus sp.WLJ097、Sphingomonas sp.WLJ118和Chryseobacterium sp.WLJ119能显著提高玉米的株高、鲜重和干重,具有植株促生能力.使用营养丰富的LB和WS培养基获得的微生物种类和特有微生物数量都更多,含nif H基因和具有植物促生能力的菌株比例更高.本研究说明沙漠植物根际蕴含了大量微生物种质资源,具有较好的开发前景.     

龙葵根际和内生Cd抗性细菌的筛选及其生物学特性

调查了南京栖霞铅锌银矿区龙葵根际土壤和植株的重金属含量,采用稀释培养法和选择性培养基分离筛选Cd抗性细菌并研究其生物学特性.结果表明,栖霞山铅锌银矿区龙葵根际土壤总Cd含量平均为19.74 mg·kg -1,污染十分严重,龙葵(Solanum nigrum)茎和叶的Cd富集系数分别为1.21 ～9.65和0.65 ～4.76,对Cd具有较强富集能力.从龙葵根际土壤和植株中分离筛选到5株Cd抗性细菌,菌株AR1和AY1分别分离自根和叶内,BGJ4和CGJ1均分离自根际土.16S rDNA序列分析表明5株Cd抗性细菌均属于芽孢杆菌属(Bacillus),能够产生吲哚乙酸(IAA)和铁载体,根内生细菌AR1在含Cd2+ 30 mg·L-1的培养基中仍能生长良好.在1.5 mg· L-1Cd2+污染条件下,Cd抗性细菌AR1、AY1、BGJ4菌株能够明显促进油菜(Oilseed rape)幼苗根的伸长,在植物-微生物联合修复重金属污染土壤方面具有潜在的应用前景.     

云南会泽废弃铅锌矿区和非矿区小花南芥根际真菌的耐镉性

采用常规、含Cd~(2+)和含Pb~(2+)的马丁氏培养基,对云南省会泽县废弃铅锌矿区和非矿区小花南芥(Arabisalpine)根际真菌进行分离,将分离的菌株接种到含不同浓度(0、0.05、0.5、5mmolL~(-1))Cd~(2+)的马铃薯葡萄糖培养液中,比较废弃铅锌矿区和非矿区小花南芥根际真菌的镉耐性.结果表明:Cd~(2+)显著抑制铅锌矿区和非矿区小花南芥根际真菌的生长;常规、含Cd~(2+)和含Pb~(2+)的马丁氏培养基分离的铅锌矿区小花南芥根际真菌Cd~(2+)的生长半数抑制浓度(EC50值)平均值和最大值均明显大于非矿区,表明铅锌矿区小花南芥根际真菌对Cd~(2+)的耐性强于非矿区;3种培养基分离的真菌中,含Cd~(2+)培养基分离的铅锌矿区小花南芥根际真菌对Cd~(2+)的耐性最强.表3参26     

活性污泥好氧异养菌群平板分离培养方法的改进

自然温度(12~21℃)、贫营养、活性污泥提取物等目前提高菌群可培养性的方法用于培养活性污泥好氧异养菌群的结果显示,这些方法均能显著提高平板培养基的分离培养能力.含活性污泥提取物的贫营养培养基ASEⅡ培养细菌的数量可占细菌总数的23.2%,在所有培养基中最高,而营养最丰富的培养基MRS培养细菌的数量只占细菌总数的8.82%,在所有培养基中最低.图1表4参14     

返青前后草地早熟禾草坪根际微生物区系动态

根际是土壤-植物生态系统物质及能量交换的活跃界面,根际微生物不仅直接影响植物对水分、养分的吸收,而且也同时影响植物对不良环境的抵抗能力。利用选择性培养基对草地早熟禾(PoapratensisL.)返青前后根际与非根际的细菌、真菌以及放线菌类群进行分离测数,拟从根际及非根际土壤微生物区系动态变化方面来阐述草地早熟禾返青前后其根际微生态的变化规律。结果表明:(1)草地早熟禾草坪的返青后,根际及非根际细菌、真菌数量明显增加,但放线菌数量呈减少趋势;(2)无论返青前后或者根际以及非根际,细菌的数量都占整个土壤微生物量的绝大部分,细菌数量的变化代表了整个微生物类群数量的变化趋势,使得草地早熟禾返青后根际土壤由"真菌型"向"细菌型"转化;(3)返青前后,各微生物类群都表现出明显的根际效应。     

废弃铅锌矿区和非矿区小花南芥根际真菌的分离及其铅耐性研究

采用常规、含Cd^2＋和含Pb^2＋马丁氏培养基,对云南省会泽县废弃铅锌矿区和非矿区小花南芥（Arabis alpina Linn）根际真菌进行分离和鉴定,将分离的菌株接种在含不同浓度（0、0．1、1和10mmol L^-1）Pb^2＋的马铃薯葡萄糖培养液中,比较废弃铅锌矿区和非矿区小花南芥根际真菌的铅耐性,结果表明：从废弃铅锌矿区和非矿区小花南芥根际共分离获得11个属的54株真菌。大于1mmolL^-1的Pb^2＋显著抑制铅锌矿区和非矿区小花南芥根际真菌的生长;含Cd^2＋和含Pb^2＋的马丁氏培养基分离的铅锌矿区小花南芥根际真菌Pb^2＋的生长半数抑制浓度（EC50）平均值和最大值均明显大于非矿区,铅锌矿区小花南芥根际真菌对Pb^2＋的耐性强于非矿区。     

几种可利用多氯联苯和多溴联苯醚植物内生菌的分离及初步鉴定

采用PDA、NA以及高氏一号培养基,分别从毛白杨、绦柳、荻以及二穗短柄草植株内分离、纯化出37株内生菌,其中34株为细菌,3株为放线菌.以分离纯化得到菌株各自对应的营养培养基并添加不同浓度的污染物对获得的37株内生菌进行污染物的耐受筛选,发现24株分别对20 mg·L-1的aroclor1254和BDE209具有一定的耐受性.利用无机盐培养基并添加污染物作为微生物唯一碳源筛选,发现菌株CPY-4和菌株SGL-1可以在只提供4-BDE的无机盐培养基中存活,菌株CPY-4,SGL-1、菌株4、菌株13可以在提供BDE209和aroclor1254的无机盐培养基中存活.形态学观察和16Sr DNA序列同源性分析,初步确定菌株SGL-1为克雷伯氏菌属,菌株CPY-4为地衣芽胞杆菌,菌株4和菌株13为肠杆菌属.     

不同微生物菌剂对当归苗生长及根际土微生物和养分的影响

通过当归拌种育苗试验研究4种微生物菌剂对当归新茬地和连作地育苗质量和品质的影响,通过测定根际土养分和可培养微生物类群数探讨添加外源微生物菌剂对当归苗根际土微生物和养分代谢功能的调节作用.结果显示:4种微生物菌剂处理在新茬地和连作地育苗条件下,均显著提高一等苗并降低废弃苗;其中复合真菌菌剂(PO+BB)处理在新茬地和连作地对有效苗数量的提高效果最好,较对照(CK)处理分别提高了6.6%和26.8%;添加外源微生物菌剂均极显著地改变根际土细菌、真菌、放线菌、氨化细菌和固氮菌等可培养微生物的类群数量(P0.001),同时也显著影响了有机质、有效钾和有效磷在根际土的积累和转化利用(P0.01).新茬地和连作地根际土微生物与养分的相关分析结果显示,连作地育苗显著改变了根际土微生物与养分的相关关系;而外源微生物菌剂处理条件下,根际土微生物与养分的相关分析结果显示,不同微生物菌剂不仅改变某类微生物与根际土养分的相关关系,而且与其他养分的积累和转化利用存在一定的协同效应.本研究表明,添加外源微生物菌剂通过影响根际微生物类群数量和功能改善了根际土养分的含量和植物的吸收利用能力,进而提高了当归苗的品质,因此在当归栽培过程中具有较好的应用前景.     

玉米内生固氮菌的回接分离及限菌条件下在玉米根内的定殖

对从玉米茎内分离的18株固氮菌进行了回接分离试验，经多碳源无氮培养基、菌落形态、颜色与REP－PCR相结合的方法，筛选、鉴定出了两株具有较高乙炔还原活性和能在玉米体内定殖的菌株R1和R6。把携带gfp标记基因的质粒pKK223-GFP和nifH-gfp的质粒pMGFP2.1分别转化R1和R6，得到携带gfp，且菌落带绿色荧光的转化子R1A和R6A，以及携带nifH-gfp的转化子R1B和R6B。在限菌条件下对R1A和R6A，在玉米根内的定殖以及R1B和R6B中nifH-gfp融合基因的表达进行了研究，结果表明了R1A和R6A主要定殖在根的皮层细胞、胞间隙、中柱细胞和导管内，有时在活的细胞内也可检测到，R1B和R6B中的融合基因因受玉米根内营养以及其它与固氮酶基因表达相关因素的限制，很难在玉米根内表达，只有在添加了1‰蔗糖后，nifH-gfp才能在根内表达。     

保水剂对侧柏容器苗根际土壤微生物种群结构及干旱适应能力的影响

为更好地将保水剂应用到干旱山地的容器苗造林中,以丙烯酰胺-丙烯酸钾交联共聚物型保水剂作为供试材料,设置育苗时(NS)和造林时(AS)各加4.0 g保水剂的两种保水剂应用方式,利用传统的平板计数法和现代的末端限制性片段长度多态性分析(Terminal restricti on fragment length polymorphism,T-RFLP)技术,研究NS和AS对侧柏容器苗造林一年后根际土壤微生物种群结构及对侧柏干旱适应能力的影响.结果表明:保水剂可显著提高侧柏根际土壤细菌、真菌的数量和微生物量碳、氮的含量,且NS处理根际土壤的细菌数量和微生物量碳分别比AS处理提高21.65%和8.04%;保水剂及其应用方式对造林初期侧柏根际土壤的细菌群落结构产生较大影响,保水剂显著增加了根际土壤中细菌群落结构的多样性,NS处理具有最高的丰富度指数、多样性指数和最低的优势度指数;此外,同对照相比,NS处理还显著提高了侧柏容器苗的造林成活率、叶片的净光合速率和相对含水量,降低了相对电导率;同AS处理相比,NS处理的造林成活率和叶片相对含水量分别提高了11.64%和8.33%,而相对电导率降低(降低12.19%).综上说明,保水剂通过改善侧柏容器苗根际土壤的微生物种群结构而提高植物的干旱适应能力,利用添加保水剂的容器苗造林具有更好的应用效果.     

油井采出液中硝酸盐还原菌的分离培养及对硫酸盐还原的抑制

采用油井采出液培养基和加入无机盐成分的改良油井采出液培养基,对大庆油田萨北过渡带油井采出液中的细菌进行分离培养及初步鉴定,比较了两种情况下培养出的具有硝酸盐和/或亚硝酸盐还原,以及/或反硝化能力菌群结构的差异.利用采出液培养基培养出一组新的微生物菌株,并且分离的硝酸盐和/或亚硝酸盐还原菌,以及/或反硝化细菌(Nitrate/nitrite reducing bacteria,denitrifying bacteria,NRDB)比例明显高于无机盐-采出液培养基;但培养基中无机盐成分的添加,提高了可培养NRDB的群落生物多样性.仅仅向油井采出液中直接投加硝酸盐作为电子受体,对其中硝酸盐还原、亚硝酸盐还原和反硝化微生物(NRDB)的激活作用以及产抑制硫化物产生的能力有限,而同时加入分离自采出液的NRDB和硝酸盐则对硫酸盐还原菌(SRB)的生长和产硫化物活性都产生了明显的抑制.但是NRDB与硝酸盐同时投加对不同SRB的抑制效果并不相同,导致了SRB群落结构的变化.图5表2参17     

培养条件对产酸克雷伯氏菌SG-11生物合成IAA影响的研究

研究了产酸克雷伯氏菌（Ｋｌｅｂｉｓｅｌｌａ ｏｘｙｔｏｃａ）ＳＧ－１１菌株的培养条件对其ＩＡＡ生物合成的影响。结果表明：该菌生长到稳定期时才开始大量产生ＩＡＡ；在基本培养基中，该菌对不同氮源的利用有别，在以（ＮＨ４）２ＳＯ４为氮源时，ＩＡＡ的产量最高，作为碳源的葡萄糖当加入浓度为６ｇ／Ｌ时，游离态ＩＡＡ的产量最高；在ＬＢ培养基中，当ｐ（Ｏ２）由正常值降低１／４时，ＩＡＡ的产生量陡然下降，灭菌前，把     

不同生存环境和磷酸盐对4株溶磷菌溶磷能力的影响

利用液体培养法研究了不同NaCl浓度、碳源、氮源、溶解氧及磷酸盐对分离自小麦(Triticum aestivum)、苜蓿(Medicago sativa)根际的4株优良溶磷细菌(Lx81、Jm92、Dm84、Lx191)溶磷能力的影响.结果表明:(1)4株溶磷菌均为高耐盐菌株,其中Jm92和Din84维持较高溶磷量的NaCl浓度最高阈值分别为4%和8%,Lx81、Lx191在NaCl浓度超过5%时溶磷活性受到抑制.(2)Lx81的最佳碳源是葡萄糖,其次为蔗糖;另外3株菌在葡萄糖和蔗糖为碳源时,均有较高的溶磷能力;4株菌都几乎不能利用淀粉.(3)Lx81和Jm92在以(NH_4)_2SO_4和NH_4 NO_3为氮源时溶磷活性均较高,(NH_4)_2SO_4 是Lx191的最佳氮源,而NH~+-N存在会导致Dm84溶磷活性降低.(4)供试4株菌均不是专性厌氧菌,但不同菌株对氧的需求量不同.(5)4株溶磷菌对Ca_3-P和Ca_8-P有较大的溶解效率,但它们几乎都不能溶解Ca_(10)-P.图3表4参14     

Nitrifying and denitrifying bacteria in aerobic granules formed in sequencing batch airlift reactors

The purpose of this study was to investigate nitrifying bacteria and denitrifying bacteria isolated from aerobic granules. Aerobic granules were formed in an internal-circulate sequencing batch airlift reactor (SBAR) and biodegradation of NH₃ ^?-N was analyzed in the reactor. Bacteria were isolated and determined from aerobic granules using selected media. The growth properties and morphology of bacteria colonies were observed by controlling aerobic or anaerobic conditions in the culture medium. It was found that bacteria in aerobic granules were diverse and some of them were facultative aerobes. The diversity of bacteria in aerobic granules was a premise of simultaneous nitrification and denitrification.     

贵州喀斯特山区不同种植年限花椒根际土壤细菌群落结构特征研究

以贵州花江峡谷花椒(Zanthoxylum bungeamun)林为研究对象,采用16S rRNA高通量测序技术,分析种植5、10、20、30 a花椒根际土壤细菌群落结构和多样性特征,探讨不同种植年限花椒土壤理化因子对根际细菌群落分布的影响,为喀斯特石漠化地区花椒农业可持续发展提供有效的理论依据。结果表明,随着花椒种植年限的增加,土壤含水率、pH和有效磷逐渐升高,有机质、铵态氮和硝态氮含量先降低后升高(P<0.05)。根际土壤细菌Shannon、Simpson指数总体呈现上升趋势,OTUs、Chao1指数在10a较低。PCo A分析显示,不同种植年限花椒根际细菌群落组成差异显著,并且随着年限的增加,群落结构趋于相似;差异指示种分析表明,5—30a差异指示种分别为Firmicutes(厚壁菌门)、Cyanobacteria(蓝藻菌门)、Planctomycetes(浮霉菌门)、Entotheonellaeota(肠杆菌门)。RDA分析表明,有机质和含水率与细菌群落分布显著相关(P<0.05)。系统发育多样性分析表明,丰富类群多样性与环境因子存在较多的正相关,稀有类群比丰富类群有更强的发育信号。Bug Base预测分析发现:随着年限增加根际土壤好氧细菌增加,厌氧细菌下降,5、10 a氧化胁迫耐受菌低于20、30 a,致病菌在5、10 a高于20、30 a。综上,种植年限影响了花椒根际土壤细菌群落多样性,不同年限根际土壤肥力不同使得土壤细菌选择性生长,随着一定时间发育花椒抗病能力提高。     

外源NO对Cu胁迫下番茄幼苗L-精氨酸代谢的影响

一氧化氮(nitric oxide,NO)作为生物活性分子,广泛参与各种生物和非生物胁迫。采用营养液培养,研究Cu胁迫下番茄体内L-精氨酸、NO、多胺代谢对外源NO的响应机制,以期为铜污染区域蔬菜种植提供科学依据和技术支持。结果表明,Cu胁迫下添加外源SNP(硝普钠,外源NO供体)能够调节番茄根系和叶片中精氨酸脱羧酶(Arginine Decarboxylase,ADC)、鸟氨酸脱羧酶(Ornithine Decarboxylase,ODC)及一氧化氮合酶(Nitric Oxide Synthase,NOS)活性。根系中,外源SNP能够上调NOS活性,L-精氨酸代谢向着NO合成方向进行;叶片中,外源SNP能够同时上调ADC、ODC、NOS活性,PA、NO的合成同时进行;此外,Cu胁迫下添加外源SNP能够增加根系和叶片L-精氨酸含量,而作为PA、NO的合成前体,精氨酸含量升高无疑会间接促进PA、NO的合成,从而提高番茄对Cu胁迫的抵抗能力。     

Multiple antibiotic resistance patterns of rhizospheric bacteria isolated from Phragmites australis growing in constructed wetland for distillery effluent treatment

Susceptibility patterns of 12 different antibiotics were investigated against rhizospheric bacteria isolated from Phragmites australis from three different zones i.e. upper (0-5 cm), middle (5-10 cm), lower (10-15 cm) in constructed wetland system with and without distillery effluent. The major pollutants of distillery effluent were phenols, sulphide, heavy metals, and higher levels of biological oxygen demand (BOD), chemical oxygen demand (COD) etc. The antibiotic resistance properties of bacteria were correlated with the heavy metal tolerance (one of distillery pollutant). Twenty-two species from contaminated and seventeen species from non-contaminated site were tested by agar disc-diffusion method. The results revealed that more than 63% of total isolates were resistance towards one or more antibiotics tested from all the three different zones of contaminated sites. The multiple-drug resistance property was shown by total 8 isolates from effluent contaminated region out of which 3 isolates were from upper zone, 3 isolates from middle zone and 2 isolates were from lower zone. Results indicated that isolates from contaminated rhizosphere were found more resistant to antibiotics than isolates from non-contaminated rhizosphere. Further this study produces evidence suggesting that tolerance to antibiotics was acquired by isolates for the adaptation and detoxification of all the pollutants present in the effluent at contaminated site. This consequently facilitated the phytoremediation of effluent, which emerges the tolerance and increases resistance to antibiotics.     

Comparison on bacterial community of rhizosphere and bulk soil of poplar plantation based on pyrosequencing北大核心CSCD

By the 454 pyrosequencing technology, this research compared the bacterial communities in poplar plantation rhizosphere and bulk soil for an accurate understanding of bacterial community colonization in the two soil environments. The species annotation showed that rhizosphere soil contained 145 bacterial genera and bulk soil contained 141 bacterial genera, with 8 common genera shared by both at a relative abundance of more than 4%. The 8 genera in common were Acidobacterium GP1, Acidobacterium GP3, Acidobacterium GP6, Gemmatimonas, Bradyrhizobium, Burkholderia, Streptomyces and Acidobacterium GP4. The relative abundance of the same bacterial community was significantly different between rhizosphere and bulk soil environments. Alpha diversity analysis showed that the bacterial community diversity of rhizosphere soil was higher than that of bulk soil, but the difference was not significant. The results of bacterial communities sorting could reflect the variation of soil bacterial communities from rhizosphere to the bulk and the spatial variation among different sampling points, indicating a contribution of about 21.2% variance of bacterial communities by the effect of rhizosphere. Beta diversity analysis showed great difference between rhizosphere and bulk soil samples in bacterial community composition. There were 15 genera specific to rhizosphere soil and 11 to bulk soil. The abundance of 23 genera, mainly cellulose degrading bacteria and nitrogen-fixing bacteria, changed significantly. Selectivity of root to rhizosphere microorganisms is an important mechanism leading to significant differences in the rhizosphere microbial community composition and structure, which may significantly impact the carbon and nitrogen cycles of the root-soil interface.     

Microbial community and functional genes in the rhizosphere of alfalfa in crude oil-contaminated soil

A rhizobox system constructed with crude oil-contaminated soil was vegetated with alfalfa (Medicago sativa L.) to evaluate the rhizosphere effects on the soil microbial population and functional structure, and to explore the potential mechanisms by which plants enhance the removal of crude oil in soil. During the 80-day experiment, 31.6% of oil was removed from the adjacent rhizosphere (AR); this value was 27% and 53%higher than the percentage of oil removed from the far rhizosphere (FR) and from the non-rhizosphere (NR), respectively. The populations of heterotrophic bacteria and hydrocarbon-degrading bacteria were higher in the AR and FR than in the NR. However, the removal rate of crude oil was positively correlated with the proportion of hydrocarbon-degrading bacteria in the rhizosphere. In total, 796, 731, and 379 functional genes were detected by microarray in the AR, FR, and NR, respectively. Higher proportions of functional genes related to carbon degradation and organic remediation, were found in rhizosphere soil compared with NR soil, suggesting that the rhizosphere selectively increased the abundance of these specific functional genes. The increase in water-holding capacity and decrease in pH as well as salinity of the soil all followed the order of AR>FR>NR. Canonical component analysis showed that salinity was the most important environmental factor influencing the microbial functional structure in the rhizosphere and that salinity was negatively correlated with the abundance of carbon and organic degradation genes.